Increased expression of LXR alpha, ABCG5, ABCG8, and SR-BI in the liver from normolipidemic, nonobese Chinese gallstone patients

Cholesterol supersaturation of bile is one prerequisite for gallstone formation. In the present study of Chinese patients with gallstones, we investigated whether this phenomenon was correlated with the hepatic expression of genes participating in the metabolism of cholesterol and bile acids. Twenty-two nonobese, normolipidemic patients (female-male, 11:11) with gallstones were investigated with 13 age- and body mass index-matched gallstone-free controls (female-male, 10:3). The bile from the gallstone patients had higher cholesterol saturation than that from the controls. The mRNA levels of ABCG5, ABCG8, and liver X receptor alpha (LXRalpha) in the gallstone patients were increased by 51, 59, and 102%, respectively, and significantly correlated with the molar percentage of biliary cholesterol and cholesterol saturation index (CSI). The mRNA and protein levels of the hepatic scavenger receptor class B type I (SR-BI) were increased, and a significant correlation was found between the protein levels and the CSI. No differences were recorded between the two groups concerning the hepatic synthesis of cholesterol, bile acids, and esterification of cholesterol. Our results suggest that the upregulation of ABCG5/ABCG8 in gallstone patients, possibly mediated by increased LXRalpha, may contribute to the cholesterol supersaturation of bile. Our data are consistent with the possibility that increased amounts of biliary cholesterol may originate from plasma HDL cholesterol by enhanced transfer via SR-BI.     

Protease-catalyzed synthesis of a precursor dipeptide, Z-Asp-Val-NH2 of thymopentin, in organic solvents

The protease-catalyzed, kinetically controlled synthesis of a precursor dipeptide, Z-Asp-Val-NH(2) of thymopentin (TP-5), in organic solvents was studied. Z-Asp-OMe and Val-NH(2) were used as the acyl donor and the nucleophile, respectively. An industrial alkaline protease alcalase was used to catalyze the synthesis of the target dipeptide in water-organic cosolvent systems. The conditions of the synthesis reaction were optimized by examining the effects of several factors, including organic solvents, water content, temperature, pH, and reaction time on the yield of Z-Asp-Val-NH(2). The optimum conditions using alcalase as the catalyst are pH 10.0, 35 degrees C, in acetonitrile/Na(2)CO(3)-NaHCO(3) buffer system (9:1, V/V), reaction time 5 h, with a yield of 63%. The dipeptide product was confirmed by LC- MS.     

间作和净作条件下喜阴植物谢君魔芋的光合作用及光合诱导特征研究

为了解种植模式对谢君魔芋(Amorphophallus xiei)光合作用的影响,研究了间作和净作模式下谢君魔芋的光合作用和光合诱导特征。结果表明,间作模式下的谢君魔芋净光合速率(Pn)比净作模式的高18.97%,最大气孔导度(gs-max)比净作模式的小22.4%,且间作模式下谢君魔芋有较高的表观量子产额(AQY)、光饱和点(LSP)、羧化效率(CE)、叶绿素含量以及较小的暗呼吸速率(Rd)、光补偿点(LCP)、CO_2补偿点(Г*)。高光照诱导后,间作模式下的光合诱导状态(IS)大于净作模式,且在间作模式下光合诱导过程中达到最大光合速率(Pmax)和最大气孔导度(gs-max)的30%、50%所需时间较短;同时,光合诱导过程中达到Pmax和gs-max的30%、50%和90%的时间(t30%P、t50%P、t90%P和t30%gs、t50%gs、t90%gs)与gs-initial呈负相关关系。因此,玉米-魔芋间作下,谢君魔芋通过增大AQY和LSP、减小Rd和LCP等来提高光的利用能力及维持碳平衡;同时通过快速的光合诱导,提高对光斑的利用能力从而增加碳获得。     

Orientation of SecA and SecB in complex, derived from disulfide cross-linking

SecA is the ATPase that acts as the motor for protein export in the general secretory, or Sec, system of Escherichia coli. The tetrameric cytoplasmic chaperone SecB binds to precursors of exported proteins before they can become stably folded and delivers them to SecA. During this delivery step, SecB binds to SecA. The complex between SecA and SecB that is maximally active in translocation contains two protomers of SecA bound to a tetramer of SecB. The aminoacyl residues on each protein that are involved in binding the other have previously been identified by site-directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy; however, that study provided no information concerning the relative orientation of the proteins within the complex. Here we used our extensive collection of single-cysteine variants of the two proteins and subjected pairwise combinations of SecA and SecB to brief oxidation to identify residues in close proximity. These data were used to generate a model for the orientation of the two proteins within the complex.     

Genome-Wide Association and Expression Analysis Revealed the Candidate Variants and Molecular Underpinnings of Cold-Stress Response in Large Yellow Croaker

Marine Biotechnology - Large yellow croaker (Larimichthys crocea) is one of the most economically important fish in China. Recently, global climate change has caused more and more intense and...     

Protein expression in accessions of Chinese onion with different allelopathic potentials under monocropping and intercropping systems

Protein expressions of Chinese onion accessions grown under monoculture and intercropped with cucumber were evaluated in pot experiments. Chinese onion accessions used were L04 (with weak allelopathic potential) and L06 (with strong allelopathic potential). Root proteins were separated by two-dimensional electrophoresis and the variable expressed protein spots were identified with MALDI-TOF-TOF mass spectrometer. Forty-seven identified proteins were classified into nine functional categories. Compared monocropping and intercropping, 31 identified variable protein spots were classified into energy metabolism (14 %), phenylpropanoid biosynthesis (28 %), organosulfur compounds biosynthesis (25 %), carbohydrate metabolism (10 %), fatty acid hydrogen peroxide metabolism (9 %), protein translation (3 %), other function (3 %), and no assigned function (9 %). Compared Chinese onion accessions of differing allelopathy potentials, 22 identified variable protein spots were classified into energy metabolism (18 %), phenylpropanoid biosynthesis (27 %), organosulfur compounds biosynthesis (23 %), carbohydrate metabolism (9 %), nucleosome component (4 %), other function (14 %), and no assigned function (5 %). The level of variable-expressed proteins involved in phenylpropanoid and organosulfur compounds biosynthesis significantly upregulated in treatments intercropped with cucumber. These results suggested that putative allelochemicals of Chinese onion were mainly produced by phenylpropanoid biosynthesis and organosulfur compounds biosynthesis pathway.     

Stable transfection of Eimeria tenella: constitutive expression of the YFP-YFP molecule throughout the life cycle

The obligate intracellular apicomplexan parasite Eimeria tenella, one of seven species of Eimeria that infect chickens, elicits protective cell-mediated immunity against challenge infection. For this reason, recombinant E. tenella parasites could be utilised as an effective vaccine vehicle for expressing foreign antigens and inducing immunity against heterologous intracellular microbes. A stable line of E. tenella expressing foreign genes is a prerequisite, and in this work an in vivo stable transfection system has been developed for this parasite using restriction enzyme-mediated integration (REMI). Two transgenic populations of E. tenella have been obtained that express YFP-YFP constitutively throughout the parasite life cycle. Southern blotting and plasmid rescue analyses show that the introduced exogenous DNA was integrated at random into the parasite genome. Although the life cycle of the transgenic populations was delayed by at least 12 h and the output of oocysts was reduced 4-fold relative to the parental BJ strain of E. tenella, the transgenic parasites were sufficiently immunogenic to protect chickens against challenge with either transgenic or parental parasites. These results are encouraging for the development of transgenic E. tenella as a vaccine vector and for more detailed investigation of the biology of the genus Eimeria.     

Kinetic investigation of the polymerase and exonuclease activities of human DNA polymerase ε holoenzyme

In eukaryotic DNA replication, DNA polymerase ε (Polε) is responsible for leading strand synthesis, whereas DNA polymerases α and δ synthesize the lagging strand. The human Polε (hPolε) holoenzyme is comprised of the catalytic p261 subunit and the noncatalytic p59, p17, and p12 small subunits. So far, the contribution of the noncatalytic subunits to hPolε function is not well understood. Using pre-steady-state kinetic methods, we established a minimal kinetic mechanism for DNA polymerization and editing catalyzed by the hPolε holoenzyme. Compared with the 140-kDa N-terminal catalytic fragment of p261 (p261N), which we kinetically characterized in our earlier studies, the presence of the p261 C-terminal domain (p261C) and the three small subunits increased the DNA binding affinity and the base substitution fidelity. Although the small subunits enhanced correct nucleotide incorporation efficiency, there was a wide range of rate constants when incorporating a correct nucleotide over a single-base mismatch. Surprisingly, the 3′→5′ exonuclease activity of the hPolε holoenzyme was significantly slower than that of p261N when editing both matched and mismatched DNA substrates. This suggests that the presence of p261C and the three small subunits regulates the 3′→5′ exonuclease activity of the hPolε holoenzyme. Together, the 3′→5′ exonuclease activity and the variable mismatch extension activity modulate the overall fidelity of the hPolε holoenzyme by up to 3 orders of magnitude. Thus, the presence of p261C and the three noncatalytic subunits optimizes the dual enzymatic activities of the catalytic p261 subunit and makes the hPolε holoenzyme an efficient and faithful replicative DNA polymerase.     

脉叶虎皮楠的生物碱成分研究

从脉叶虎皮楠(Daphniphyllum paxianum)的枝干中分离并鉴定了四个生物碱,分别为daphnilactone A(1)、daphnicyclidin D(2)、daphniphylline(3)、daphnicyclidin H(4).其中首次对化合物1的碳谱数据进行了归属.     

拟南芥花粉细胞质游离钙离子荧光测定法

以拟南芥花粉为材料,利用低温装载法在完整的花粉粒中,成功地载入酯化形式的钙离子荧光探针Fura-2/AM。利用荧光比率分析法对花粉细胞质中游离钙离子的分布特点进行研究并测定了花粉细胞内游离钙离子浓度。结果表明花粉萌发初期细胞质内游离钙离子呈极性分布,萌发沟附近的钙离子浓度明显高于其它部位,萌发孔附近最高,花粉细胞核中最低。花粉粒细胞萌发状态下的[Ca2 ]i=246±38nmol/L,该值与花粉粒细胞萌发状态下游离钙离子浓度用其它方法测得值接近,进一步表明所建立的用Fura-2/AM检测拟南芥花粉粒细胞质游离钙离子的方法是可靠的。